Effects of repeated lengthening contractions on skeletal muscle adaptations in female rats.

We examined the adaptation of plantar flexor muscles of female rats to 6 weeks (5 days/week) of lengthening contractions. After repeated lengthening contractions, a decrease in myofiber area of gastrocnemius medialis (26%) was accompanied by an increase in extracellular matrix (ECM) (42%) and collagen content (30.9%) without changes in muscle mass. Decrease in myofiber area (13%) and muscle mass of soleus (19%) was associated with increased collagen content (28%) and ECM (15%). Relative number of soleus myofibers stained for fast myosin increased by 26%. For plantaris, increases in collagen content (32.3%), percent ECM (17%), and myofiber area (6%) were recorded. We also observed (1) increases (3.3%) in the collagen content of the Achilles tendon, (2) no change in the crosslink content of any of the tissues tested, and (3) no difference in the force-frequency relationship of the plantar flexor muscles. Substantial decreases in myofiber areas with increases in muscle connective tissue by 6 weeks of repeated lengthening contractions did not appear to result in isometric force loss.

Impact of repetition number on muscle performance and histological response.

UNLABELLED: Skeletal muscle injury is major concern in sport- and occupation-related fields. PURPOSE: We investigated the effects of increasing stretch-shortening contraction (SSC) repetition number in vivo and the resulting changes in functional performance and quantitative morphometry in rat skeletal muscle. METHODS: Functional testing was performed on the ankle dorsiflexor muscles of Sprague-Dawley rats, which were randomly exposed to 30 SSC, 70 SSC, 150 SSC, or 15 isometric contractions of equal duration. Changes in functional performance and muscle morphometry were assessed at 48 h after exposure. Stereology was used to quantify the volume density of degenerative myofibers and normal myofibers in the tibialis anterior muscle from each group, as well as measures of inflammation and swelling and changes in the interstitial space. RESULTS: At 48 h there was a significant decline in isometric force for the 70- and 150-SSC groups (P < 0.05 and P < 0.05, respectively). Stereological measures indicated significant decreases in the percentage of volume density of normal myofibers in the 70- and 150-SSC groups (P < 0.05). Measures for percentage of volume density of degenerative myofibers and inflammation were increased (P < 0.0001 and P < 0.05, respectively) in the 70- and 150-SSC groups. Moreover, a significant increase in the percentage of volume density of degenerative myofibers in the 150-SSC group compared with the 70-SSC group was observed (P < 0.05). CONCLUSION: These data strongly suggest that exposure to increasing SSC repetitions results in increased functional decrements and morphometric indices of myofiber degeneration and inflammation, and that there is an apparent threshold (repetition number) at which this occurs.

Chronic exposure to stretch-shortening contractions results in skeletal muscle adaptation in young rats and maladaptation in old rats.

The objective of this research was to investigate skeletal muscle response to a chronic administration of stretch-shortening cycles (SSCs) in young and old rats. Dorsiflexor muscles of old (30 months, n = 5) and young (12 weeks, n = 6) rats were exposed 3 times/week for 4.5 weeks to a protocol of 80 maximal SSCs per exposure in vivo. Skeletal muscle response was characterized by isometric and dynamic performance, as well as by muscle wet mass and quantitative morphological analyses following the exposure period. The performance of the young and old groups was not statistically different at the start of the exposure. By the end of the exposure, however, a statistical difference was noted, as performance increased significantly in the young animals and decreased significantly in the old animals. Muscle wet mass of the left tibialis anterior (TA) in the treated limb was significantly greater in the youngthan in the old animals (p < 0.001), whereas there was no difference in the contra-lateral TA. No degenerative myofibers or changes in non-cellular interstitium were noted in either age group, but a significant increase was observed in the volume of the cellular interstitium in the exposed limb of the old animals (p = 0.01), which is indicative of an inflammatory response. Thus, a chronic exposure of SSCs results in significant performance increase and muscle hypertrophy in young animals, and a significant performance decrease and an increased cellular interstitial response in old animals. These findings suggest that age may impair the ability of skeletal muscle to adapt to repetitive mechanical loading, even in the absence of degeneration.

Quantitative histology and MGF gene expression in rats following SSC exercise in vivo.

PURPOSE: We investigated the effects of muscle length during stretch-shortening cycles (SSC) in vivo on changes in MGF gene expression and quantitative morphometry in rat skeletal muscle. METHODS: Dorsiflexor muscles of male Sprague-Dawley rats were exposed to seven sets of 10 SSC at 500 degrees .s(-1). Animals were randomly assigned to a long muscle length injury group (L-inj), short muscle length injury group (S-inj), or isometric group (Iso), with recoveries examined at 6 or 48 h post-injury for each group. Following exposure, animals were euthanized, and the tissue was prepared for either histology (quantitative morphometry) or RNA isolation, followed by quantitative real-time reverse transcriptase polymerase chain reaction. mRNA levels were measured for mechano-growth factor (MGF), while 18S ribosomal RNA served as the internal reference sample. RESULTS: Stereological measures indicative of edema and myofiber degeneration were significantly increased in the L-inj SSC group at 48 h when compared with the S-inj or Iso group. MGF mRNA was increased transiently at 6 h in the isometric group. In contrast, MGF mRNA was increased at 48 h in the S-inj, but was not increased at either time point in the L-inj group. CONCLUSION: These data strongly indicate that exposure to SSC at longer muscle lengths result in greater morphometric indices of inflammation and degeneration than SSC conducted at a shorter muscle lengths or isometric contractions, at the same time that the adaptation to SSC was prolonged and, apparently, not resolved in the L-inj group that was manifested by the lack of up-regulation in MGF mRNA.

Comparison of mechanical properties of rat tibialis anterior tendon evaluated using two different approaches.

Tendon injuries may result in variations of its mechanical properties. The published data of the tendon stiffness of small animals, such as mouse and rat, are exclusively obtained by measuring grip-to-grip (g-t-g) displacement. Local strain concentration and relative sliding of the specimens in the clamps might significantly affect the measured tendon deformation. In the present study, the mechanical properties of the rat tibialis anterior tendon measured using the proposed tendon mark method were compared to those evaluated using the g-t-g displacement method. Five male Sprague Dawley rats ( approximately 418 g) were used in this study. For the proposed method, reference marks were made on the tendons using permanent ink. A microscope video system was customized to observe and record the tendon deformation. Pattern recognition software was developed to obtain the displacement time-histories of the reference marks. The distance between the grips was approximately 7 mm; and the distance between the reference marks used for the data processing was approximately 5 mm. The cross-section areas of the specimens were measured using a custom-made slot gauge and by applying a constant compressive stress (0.15 MPa). The tendons were clamped between two custom-made metal grips and stretched on a testing machine at a constant speed (1 mm/s) up to failure. Throughout the tests, the tendon specimens were submerged in a PBS bath at 22 degrees C. The deformation of the specimens was evaluated using the g-t-g displacement method and the proposed method. The stress/strain curves obtained by using the g-t-g displacement can be characterized by an initial toe zone, a quasi-linear zone, and a final failure stage. The stress/strain curves determined using the proposed method are quite different from those obtained using the g-t-g displacement: it has a smaller toe zone and a stress-hardening transition, over which the tendon stiffness increases dramatically with the increasing strain. The tendon stiffness measured by using the g-t-g displacement method may underestimate the actual mechanical properties of tendon by approximately 43%.